Method for analyzing nucleobases on a single molecular basis
a nucleobase and molecular basis technology, applied in the field of single molecular basis nucleobase analysis, can solve the problems of long analysis period, achieve the effect of facilitating electron tunneling in stm between a nucleobase and its complementary nucleobase molecular tip, and facilitating electron tunneling through the complementary base-pair
Inactive Publication Date: 2012-02-28
JAPAN SCI & TECH CORP
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Benefits of technology
The present invention is related to a method for analyzing nucleobases on a single molecular basis using scanning tunneling microscopy. The inventors found that electron tunneling between a nucleobase and its complementary nucleobase molecular tip was much facilitated compared to non-complementary counterparts, allowing for selective discrimination of each nucleobase from the others. The first invention is a method for pinpointing a target nucleobase in a nucleic acid by scanning a molecular tip on the nucleobase and measuring the tunneling current between the nucleobase and the molecular tip. The second invention is a molecular tip for scanning tunneling microscopy, which is a metal tip chemically modified with adenine, guanine, cytosine, thymine, or uracil. The third invention is a set of four molecular tips for scanning tunneling microscopy, which consists of four metal tips chemically modified with adenine, guanine, cytosine, or thymine. The invention allows for the analysis of nucleobases on a single molecular basis, which can be useful in gene engineering and molecular biology.
Problems solved by technology
Therefore, establishment of a method for detecting and analyzing a specific gene sequence or a mutant sequence is an important challenge in various fields such as medical science, medical jurisprudence, molecular biology and so on.
However, these methods have some problems such as a prolonged period of time for analysis.
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[0066]The present inventions will be described in detail by the following Examples, but the present inventions are not limited to the Examples.
1. Materials and Methods
Preparation of Thiol Derivatives of Adenine, Guanine, Cytosine, and Uracil
[0067]As sample thiol derivatives of nucleobase (FIG. 1b), 4-amino-8-mercaptopyrmidine (1) (reference 19) and 2,4-hydroxy-6-mercaptopyridine (4) (reference 20) were synthesized, and the products were characterized by 1H NMR, 13C NMR, and elemental analysis. 4-Amino-2-hydoxy-6-mercaptopyrmidine (3: Sigma-Aldrich library of rare chemicals) and 2-amino-6-hydroxy-8-mercaptopurine (2: Acros) were recrystallized three times from ethanol prior to use. 2-Mercaptobenzimidazole (98%, Aldrich) and thiophenol (95%, Wako Pure Chemical, Osaka) were used without further purification. Instead of a thymine thiol derivative, a uracil derivative (4) was used because of its ease for synthesis.
[0068]1 (thiol derivative of adenine): 1H NMR (500 MHz, DMSO-d6) δ 6.74 (s...
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Abstract
A method is herein presented for analyzing nucleobases on a single molecular basis (a single molecule detection), which comprises scanning a molecular tip chemically modified with the complementary nucleobase on the nucleobases, and measuring the tunneling currents between the scanned nucleobases and the molecular tips with scanning tunneling microscopy.
Description
[0001]This application is a U.S. national stage of International Application No. PCT / JP2005 / 024285 filed Dec. 28, 2005.TECHNICAL FIELD[0002]The present invention relates to a method for analyzing nucleobases on a single molecular basis (a single molecule detection). More specifically, the present invention relates to a method for electrically pinpointing or typing a nucleobase in a nucleic acid, or electrically sequencing a nucleic acid by using scanning tunneling microscopy.BACKGROUND ART[0003]Recently, genomic structures of various species are being clarified due to technical development in molecular biology, and involvement of gene mutations have been revealed in many genetic diseases and viral diseases. Therefore, establishment of a method for detecting and analyzing a specific gene sequence or a mutant sequence is an important challenge in various fields such as medical science, medical jurisprudence, molecular biology and so on.[0004]Among the gene mutations, attention is focu...
Claims
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IPC IPC(8): G01N33/00C07D473/00C12Q1/68G01Q60/10G01Q60/16G01Q80/00
CPCB82Y35/00C12Q1/6816C12Q1/6869G01Q60/16G01Q70/18B82Y15/00Y10T436/143333C12Q2565/601
Inventor UMEZAWA, YOSHIOOHSHIRO, TAKAHITO
Owner JAPAN SCI & TECH CORP